Critical evaluation of a theory of molecular recognition using human insulin-like-growth-factor-I fragment 21–40 and its complementary peptide

“…Numbers in brackets following each sequence refer to its location within the sequence of rbGH. Peptides were synthesised by standard solid-phase fluorenylmethoxycarbonyl chemistry [17] and analysed by reverse-phase HPLC as described previously [18]. A control peptide (CRSPELETFSC) was synthesied by solid-phase fluorenylmethoxycarbonyl chemistry on an Applied Biosystems 431A automated peptide synthesiser and was analysed as described above.…”

Section: Animalsmentioning

confidence: 99%

The use of multiple‐pin peptide synthesis in an analysis of the continuous epitopes recognised by various anti‐(recombinant bovine growth hormone) sera

Beattie

Fawcett

Flint

1992

European Journal of Biochemistry

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A recently developed technology called epitope scanning permits the rapid and accurate delineation of continuous stretches of amino acids in a protein which constitute the sequential epitopes recognised by an antiserum raised to that protein. In the present report, we describe the use of this technique to identify the epitopes in the recombinant bovine growth-hormone (rbGH) molecule recognised by three polyclonal guinea-pig antisera and two polyclonal rabbit antisera. The results obtained show that, for guinea-pig antisera, 3 or 4, very-well-defined major continuous epitopes are present. As would be expected given the intrinsic genetic factors (major histocompatibility restriction, antigen processing and presentation) controlling the immune response in individual animals, subtle differences are evident in the precise location and relative reactivities of these epitopes in different guinea-pig antisera. Nevertheless, there is a large degree of overlap in these epitopes, such that immunodominant regions of the antigen can be clearly delineated. In a structural sense, these epitopes share a common motif in that they are sited in areas of the protein antigen with little secondary structure (loop/coil), although there is some contribution by neighbouring a-helices. For the two rabbit antisera, the response tends to be rather more heterogeneous, with recognition of more peptides and less clearly defined epitopes than was the case with the guinea-pig antiserum.Comparison of the four guinea-pig epitopes, identified by our experimental methods with computer predictions for this molecule (Jameson-Wolf antigenic index), indicate that two are strongly predicted, one is weakly predicted and one is not predicted. These observations, together with the displayed intraspecies and interspecies variation clearly indicate the limitations of these predictive methods.In conclusion, we have demonstrated that, despite the expected variation in the exact location of continuous epitopes defined by different anti-rbGH sera, there are large regions of overlap defining immunogenic core regions within the molecule. We believe that studies of this nature, together with further understanding of antigen processing and peptide presentation to immune cells, may have a role to play in the development of candidate peptide vaccines.Classically, the binding sites on protein antigens (epitopes) recognised by antibodies have been categorised as either continuous or discontinuous. Continuous epitopes are defined by amino acid residues which are sequential in the sequence of the polypeptide antigen, whereas discontinuous epitopes are formed by residues proximate to each other in the threedimensional structure of the protein, although not necessarily so in the sequence [l].Recently, however, this definition has been challenged on both theoretical [2] and experimental [3] grounds. For example, studies which have examined the cocrystallisation of protein antigen with antibody have found all epitopes on the antigen to be discontinuous (or conformational) in ...

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“…The same research group later found a similar highaffinity association between the g-endorphin peptide and a synthetic 5' 33' complementary peptide whose sequence had been deduced in the same way. [54] There is, as yet, no clear explanation as to why specific sense/complementary peptide interactions should not be observed reproducibly in these experiments whilst being seen with such apparent ease in others. Some of these applications will be discussed in a little more detail below.…”

Section: Observations With Sense/complementary Peptide Systemsmentioning

confidence: 74%

Specific Interactions Between Sense and Complementary Peptides: The Basis for the Proteomic Code

et al. 2002

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Critical evaluation of a theory of molecular recognition using human insulin-like-growth-factor-I fragment 21–40 and its complementary peptide

Cited by 9 publications

References 38 publications

Prediction of potential protein‐protein interaction sites from amino acid sequence

Prediction of potential protein‐protein interaction sites from amino acid sequence

The use of multiple‐pin peptide synthesis in an analysis of the continuous epitopes recognised by various anti‐(recombinant bovine growth hormone) sera

Specific Interactions Between Sense and Complementary Peptides: The Basis for the Proteomic Code

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